Novel living cell sheet harvest system composed of thermoreversible methylcellulose hydrogels

Chun Hung Chen, Chen Chi Tsai, Wannhsin Chen, Fwu Long Mi, Hsiang Fa Liang, Sung Ching Chen, Hsing Wen Sung

研究成果: 雜誌貢獻文章同行評審

97 引文 斯高帕斯(Scopus)


In this study, a novel yet simple method, using a thermoreversible hydrogel system coated on tissue culture polystyrene (TCPS) dishes, was developed for harvesting living cell sheets. The hydrogel system was prepared by simply pouring aqueous methylcellulose (MC) solutions blended with distinct salts on TCPS dishes at 20 °C. For the applications to cell culture, only those aqueous MC compositions that may form a gel at 37 °C were chosen for the study. It was found that the hydrogel coating composed of 8% MC blended with 10 g/L PBS (phosphate buffered saline) (the MC/PBS hydrogel, with a gelation temperature of ∼25 °C) stayed intact throughout the entire course of cell culture. To improve cell attachments, the MC/PBS hydrogel at 37 °C was evenly spread with a neutral aqueous collagen at 4 °C. The spread aqueous collagen gradually reconstituted with time and thus formed a thin layer of collagen (the MC/PBS/collagen hydrogel). After cells reached confluence, a continuous monolayer cell sheet formed on the surface of the MC/PBS/collagen hydrogel. When the grown cell sheet was placed outside of the incubator at 20 °C, it detached gradually from the surface of the thermoreversible hydrogel spontaneously, without treating with any enzymes. The results obtained in the MTT assay demonstrated that the cells cultured on the surface of the MC/PBS/collagen hydrogel had an even better activity than those cultured on an uncoated TCPS dish. After harvesting the detached cell sheet, the remaining viscous hydrogel system is reusable. Additionally, the developed hydrogel system can be used for culturing a multilayer cell sheet. The obtained living cell sheets may be used for tissue reconstructions.

頁(從 - 到)736-743
出版狀態已發佈 - 三月 2006

ASJC Scopus subject areas

  • 有機化學
  • 生物化學、遺傳與分子生物學 (全部)
  • 聚合物和塑料
  • 材料化學


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